| Project/Area Number |
06660061
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
植物保護
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| Research Institution | Biotechnology Institute, Akita Prefectural College of Agriculture |
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Principal Investigator |
KITAGAWA Yoshichika Biotechnology Institute, Akita Prefectural College of Agriculture, 附属生物工学研究所, 教授 (10006752)
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| Project Period (FY) |
1994 – 1995
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| Project Status |
Completed (Fiscal Year 1995)
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| Budget Amount *help |
¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 1995: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1994: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | Single chain antibody / double-stranded RNA / virus disease / variable region of antibody / モノクローン抗体 / 抗体遺伝子 / 核酸 |
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| Research Abstract |
The variable regions, VH and VL,of antibody gene against double-stranded RNA were cloned in vector in 1995. These genes were amplified with the primers of commercial kit by PCR.Single chain antibody gene (ScFv) constructed with these VH and VL transformed into bacteria ; however, the gene products did not show any binding activity to double-stranded RNA.In 1996 new VH and VL fragments were amplified with different primers from mRNA of antibody producing hybridoma cells by PCR and analyzed sequence. Sequence of these VH and VL differ from the VH and VL previously isolated in one base in 3'-terminal. We retried to construct the ScFv gene with new VH,VL and linker sequence and expressed in bacteria ; however, no binding activity were detected in gene products. At present it is still unclear whether the linker sequence is not optimal or ScFv antibody can not recognize double-stranded RNA.
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